More exploration on medicinal plants and other natural products in the present era of increase in poverty level and multi-drug resistance has become crucial. The aim of this study is to explore the inhibitory activities of Vitellaria paradoxa seed oil extract on isolated staphylococcal conjunctivitis. Cultured sample of Staphylococcus aureus isolated from a patient’s eye discharge in the Teaching Hospital Laboratory of the Imo State University, Nigeria having been diagnosed with bacterial conjunctivitis at the eye Clinic. After the incubation period, the diameter of zones of inhibition both horizontal and vertical were measured. Concentrations (100, 50 and 25mg/ml) of the ethanolic seed oil extract of V. paradoxa were assayed for the antibacterial activity - Minimum Inhibitory Concentration (MIC) using the agar well diffusion method. Ethanolic seed oil extract of V. paradoxa at concentration of 100mg /ml exhibited the highest zone of inhibition at 37.4mm for 24hrs followed by 50mg /ml and lowest using 25mg/ml (5.0mm) indicating a concentration-dependent inhibitory effect on Staphylococcal conjunctivitis. S. aureus isolated from conjunctivitis swab was susceptible to ethanolic seed oil extract of V. paradoxa at 100mg/ml, 50mg/ml and 25mg/ml concentrations, suggesting ethanolic extract of V. paradoxa oil as possessing antimicrobial property. Further exploration for its use as an ocular anti-bacterial agent is recommended.